Cancerous B cell treatment using substituted nucleoside derivatives

Abstract

Processes for the killing of cancerous B cells, and particularly chronic lymphocytic leukemia (CLL) cells are disclosed. In one process, cancerous B cells that do not proliferate when contacted with an immune response-enhancing agent are contacted with an amount of such an agent sufficient to cause peripheral CLL cells to undergo blast transformation and proliferation. The contacted cells are then maintained for a time period sufficient for them to die from that contact. Further contacting of those cells with a cytotoxic amount of an anti-cancer drug or cytotoxic conjugate enhances the death of those cancer cells. In another process, peripheral CLL cells that proliferate on contact with an immune response-enhancing-agent are contacted with a proliferation-inducing amount of such an agent. The contacted cells are maintained for a time period sufficient to undergo blast transformation and proliferation, and the blasts are then contacted with a cytotoxic amount of an anti-cancer drug or cytotoxic conjugate and maintained.

What is claimed is: 
     
       1. A process for killing cancerous B cells that comprises: (a) contacting in an aqueous medium a CLL cell proliferation-inducing amount of an immune response-enhancing agent with cancerous B cells of a host mammal that do not undergo blast transformation and proliferation when contacted with an immune response-enhancing agent; and   (b) maintaining said contact under biological culture conditions for a time period sufficient for the contacted cancerous B cells to die;   said immune response-enhancing agent having a structure that corresponds to a formula ##STR12## wherein Z is O, S or N--R 2  ;   R 1  contains up to about 20 atoms and has a Hammett substituent sigma constant for ionization of a meta-substituted benzoic acid that is greater than that of hydrogen;   R 2  is a radical having a length up to about that of an n-decyl group that is selected from the group consisting of C 1  -C 10  alkyl, C 3  -C 10  beta-alkenyl, phenyl-substituted C 3  -C 6  beta-alkenyl, benzyl, C 1  -C 6  alkoxybenzyl, nitrobenzyl, hydroxy C 1  -C 10  alkyl, polyhydroxy C 1  -C 10  alkyl, halo C 1  -C 10  alkyl, polyhalo C 1  -C 10  alkyl, C 1  -C 6  alkylene C 1  -C 6  alkylcarboxylate, C 1  -C 10  alkanoyl, C 1  -C 6  alkoxy C 1  -C 6  alkylenecarbonyl, and C 1  -C 6   alkylenecarboxamido in which the carboxamido group has the formula CONR 9  R 10  wherein R 9  and R 10  are the same or different and are selected from the group consisting of hydrogen and C 1  -C 6  alkyl or NR 9  R 10  together form a heterocyclic ring having five or six atoms in the ring;   X is oxygen or sulfur;   R 3  is a radical selected from the group consisting of hydrogen, C 1  -C 6  alkyl, hydroxy C 1  -C 6  alkyl, polyhydroxy C 1  -C 6  alkyl, phenyl, phenyl-C 1  -C 6  alkylene, C 1  -C 6  alkylphenyl, C 1  -C 6  alkoxyphenyl, halophenyl, trifluoromethylphenyl, hydroxy, oxo (O═), C 1  -C 6  alkoxy, phenyl-C 1  -C 6  alkoxy, halo, mercapto, thioxo (S═), C 1  -C 6  alkylthio, phenyl-C 1  -C 6  alkylthio, C 1  -C 6  alkanoyl (C 1  -C 6  acyl), C 1  -C 6  alkoxy carbonyl, C 1  -C 6  alkylene C 1  -C 6  alkylcarboxylate, C 1  -C 6  alkoxy C 1  -C 6  alkyl carbonyl, and C 1  -C 6  alkyl carboxamido in which the carboxamido group has the formula CONR 9  R 10  wherein R 9  and R 10  are the same or different and are selected from the group consisting of hydrogen and C 1  -C 6   alkyl, or NR 9  R 10  together form a heterocyclic ring having five or six atoms in the ring;   R 4  is a beta-bonded aldoglycoside radical selected from the group consisting of 1'-aldopentosidyl, 1'-aldohexosidyl, mono-deoxygenated 1'-aldopentosidyl, and mono-deoxygenated 1'-aldohexosidyl, and their O-substituted C 1  -C 6  alkyl, C 1  -C 6  alkanoyl, benzyl, benzoyl and C 1  -C 6  acetal or ketal derivatives, an O-substituent other than an acetal or ketal, if present on one oxygen, being present on all available ring substituent oxygens;   the pharmaceutically acceptable salts of said agent; and   the tautomers thereof,   said immune response-enhancing agent being free from ionic charge in water at pH 7.2-7.4.   
     
     
       2. The process according to claim 1 wherein said immune response-enhancing agent has a structure that corresponds to a formula ##STR13## wherein Z is N--R 2  and ═X is ═O. 
     
     
       3. The process according to claim 2 wherein R 4  is 1'-aldopentosidyl in mono-deoxygenated 1'-aldopentosidyl. 
     
     
       4. The process according to claim 3 wherein R 1  is OH or SH. 
     
     
       5. The process according to claim 3 wherein R 2  is selected from the group consisting of C 1  -C 10  alkyl, C 3  -C 10  beta-alkenyl and halo C 1  -C 10  alkyl. 
     
     
       6. The process according to claim 1 wherein said immune response-enhancing agent is 7-allyl-8-oxoguanosine, 7-(1-chloroethyl)-8-oxoguanosine or 8-mercaptoguanosine. 
     
     
       7. The process according to claim 1 wherein said contacting is carried out in vitro in a mammalian cell culture medium. 
     
     
       8. The process according to claim 7 wherein said mammalian cell culture medium includes about 5 to about 15 volume percent fetal calf serum or autologous plasma. 
     
     
       9. The process according to claim 1 wherein said cancerous B cells are circulating chronic lymphocytic leukemia, hairy cell leukemia cells or non-Hodgkins' leukemia cells. 
     
     
       10. The process according to claim 1 including the further steps of contacting the maintained cells of step (b) with a cytotoxic amount of an anti-cancer drug or a two portion conjugate molecule, one portion binding to a cell surface antigen that is expressed in enhanced amounts due to said contacting of step (a) and the other portion being an anti-cancer drug or a cytoxic agent, said further contacting being at a time about 1 to about 4 days after the first contacting of step (a), and maintaining the anti-cancer drug-contacted cells under biological culture conditions for a time period sufficient for those cells to die. 
     
     
       11. The process according to claim 10 wherein said immune response-enhancing agent has a structure that corresponds to a formula ##STR14## wherein Z is N--R 2  and ═X is ═O. 
     
     
       12. The process according to claim 11 wherein R 4  is 1'-aldopentosidyl in mono-deoxygenated 1'-aldopentosidyl. 
     
     
       13. The process according to claim 12 wherein R 1  is OH or SH. 
     
     
       14. The process according to claim 12 wherein R 2  is selected from the group consisting of C 1  -C 10  alkyl, C 3  -C 10  beta-alkenyl and halo C 1  -C 10  alkyl. 
     
     
       15. The process according to claim 10 wherein said immune response-enhancing agent is 7-allyl-8-oxoguanosine, 7-(1-chloroethyl)-8-oxoguanosine or 8-mercaptoguanosine. 
     
     
       16. The process according to claim 10 wherein said cancerous B cells are circulating chronic lymphocytic leukemia, hairy cell leukemia cells or non-Hodgkins' leukemia cells. 
     
     
       17. The process according to claim 10 wherein said anti-cancer drug is selected from the group consisting of etoposide, cytoxan, adriamycin, vincristine, cisplatin, chlorambucil, methotrexate, carmustine, cytarabine, dexamethasone and doxorubicin. 
     
     
       18. A process for killing chronic lymphocytic leukemia (CLL) cells that comprises: (a) contacting in an aqueous medium a proliferation-inducing amount of an immune response-enhancing agent with human CLL cells that undergo blast transformation and proliferation when contacted with said immune response-enhancing agent with a proliferation-inducing amount of an immune response-enhancing agent;   (b) maintaining said contact under biological culture conditions for a time period sufficient for said contacted CLL cells to proliferate and form blasts;   (c) contacting said blasts with a cytotoxic amount of an anti-cancer drug; and   (d) maintaining said contact with said anti-cancer drug under biological culture conditions for a time period for said contacted blast cells to die,   said immune response-enhancing agent having a structure that corresponds to a formula ##STR15## wherein Z is O, S or N--R 2  ;   R 1  contains up to about 20 atoms and has a Hammett substituent sigma constant for ionization of a meta-substituted benzoic acid that is greater than that of hydrogen;   R 2  is a radical having a length up to about that of an n-decyl group that is selected from the group consisting of C 1  -C 10  alkyl, C 3  -C 10  beta-alkenyl, phenyl-substituted C 3  -C 6  beta-alkenyl, benzyl, C 1  -C 6  alkoxybenzyl, nitrobenzyl, hydroxy C 1  -C 10  alkyl, polyhydroxy C 1  -C 10  alkyl, halo C 1  -C 10  alkyl, polyhalo C 1  -C 10  alkyl, C 1  -C 6  alkylene C 1  -C 6  alkylcarboxylate, C 1  -C 10  alkanoyl, C 1  -C 6  alkoxy C 1  -C 6  alkylenecarbonyl, and C 1  -C 6  alkylenecarboxamido in which the carboxamido group has the formula CONR 9  R 10  wherein R 9  and R 10  are the same or different and are selected from the group consisting of hydrogen and C 1  -C 6  alkyl, or NR 9  R 10  together form a heterocyclic ring having five or six atoms in the ring;   X is oxygen or sulfur;   R 3  is a radical selected from the group consisting of hydrogen, C 1  -C 6  alkyl, hydroxy C 1  -C 6  alkyl, polyhydroxy C 1  -C 6  alkyl, phenyl, phenyl-C 1  -C 6  alkylene, C 1  C 6  alkylphenyl, C 1  -C 6  alkoxyphenyl, halophenyl, trifluoromethylphenyl, hydroxy, oxo (O═), C 1  -C 6  alkoxy, phenyl-C 1  -C 6  alkoxy, halo, mercapto, thioxo (S═), C 1  -C 6  alkylthio, phenyl-C 1  -C 6  alkylthio, C 1  -C 6  alkanoyl (C 1  -C 6  acyl), C 1  -C 6  alkoxy carbonyl, C 1  -C 6  alkylene C 1  -C 6  alkylcarboxylate, C 1  -C 6  alkoxy C 1  -C 6  alkyl carbonyl, and C 1  -C 6  alkyl carboxamido in which the carboxamido group has the formula CONR 9  R 10  wherein R 9  and R 10  are the same or different and are selected from the group consisting of hydrogen and C 1  -C 6  alkyl or NR 9  R 10  together form a heterocyclic ring having five or six atoms in the ring;   R 4  is a beta-bonded aldoglycoside radical selected from the group consisting of 1'-aldopentosidyl, 1'-aldohexosidyl, mono-deoxygenated 1'-aldopentosidyl, and mono-deoxygenated 1'-aldohexosidyl, and their O-substituted C 1  -C 6  alkyl, C 1  -C 6  alkanoyl, benzyl, benzoyl and C 1  -C 6  acetal or ketal derivatives, an O-substituent other than an acetal or ketal, if present on one oxygen, being present on all available ring substituent oxygens;   the pharmaceutically acceptable salts of said agent; and   the tautomers thereof,   said immune response-enhancing agent being free from ionic charge in water at pH 7.2-7.4.   
     
     
       19. The process according to claim 18 wherein said immune response-enhancing agent has a structure that corresponds to a formula ##STR16## wherein Z is N-R 2  and ═X is ═O. 
     
     
       20. The process according to claim 19 wherein R 4  is 1'-aldopentosidyl in mono-deoxygenated 1'-aldopentosidyl. 
     
     
       21. The process according to claim 20 wherein R 1  is OH or SH. 
     
     
       22. The process according to claim 20 wherein R 2  is selected from the group consisting of C 1  -C 10  alkyl, C 3  -C 10  beta-alkenyl and halo C 1  -C 10  alkyl. 
     
     
       23. The process according to claim 18 wherein said immune response-enhancing agent is 7-allyl-8-oxoguanosine, 7-(1-chloroethyl)-8-oxoguanosine or 8-mercaptoguanosine. 
     
     
       24. The process according to claim 18 wherein said contacting is carried out in vitro in a mammalian cell culture medium. 
     
     
       25. The process according to claim 24 wherein said mammalian cell culture medium includes about 5 to about 15 volume percent fetal calf serum or autologous plasma. 
     
     
       26. The process according to claim 18 including the further steps of contacting the maintained cells of step (b) with a cytotoxic amount of an anti-cancer drug or a two portion conjugate molecule, one portion binding to a cell surface antigen that is expressed in enhanced amounts due to said contacting of step (a) and the other portion being an anti-cancer drug or a cytoxic agent, said further contacting being at a time about 1 to about 4 days after the first contacting of step (a), and maintaining the anti-cancer drug-contacted cells under biological culture conditions for a time period sufficient for those cells to die. 
     
     
       27. A process for killing cancerous B cells that comprises: (a) contacting in an aqueous medium a CLL cell proliferation-inducing amount of an immune response-enhancing agent with cancerous B cells of a host mammal that undergo blast transformation and proliferation when contacted with an immune response-enhancing agent;   (b) maintaining said contact under biological culture conditions for a time period sufficient for said contacted cancerous B cells to proliferate and form blasts, and for the contacted cancerous B cells to die;   said immune response-enhancing agent having a structure that corresponds to a formula ##STR17## wherein Z is O, S or N--R 2  ;   R 1  contains up to about 20 atoms and has a Hammett substituent sigma constant for ionization of a meta-substituted benzoic acid that is greater than that of hydrogen;   R 2  is a radical having a length up to about that of an n-decyl group that is selected from the group consisting of C 1  -C 10  alkyl, C 3  -C 10  beta-alkenyl, phenyl-substituted C 3  -C 6  beta-alkenyl, benzyl, C 1  -C 6  alkoxybenzyl, nitrobenzyl, hydroxy C 1  -C 10  alkyl, polyhydroxy C 1  -C 10  alkyl, halo C 1  -C 10  alkyl, polyhalo C 1  -C 10  alkyl, C 1  -C 6  alkylene C 1  -C 6  alkylcarboxylate, C 1  -C 10  alkanoyl, C 1  -C 6  alkoxy C 1  -C 6  alkylenecarbonyl, and C 1  -C 6  alkylenecarboxamido in which the carboxamido group has the formula CONR 9  R 10  wherein R 9  and R 10  are the same or different and are selected from the group consisting of hydrogen and C 1  -C 6  alkyl or NR 9  R 10  together form a heterocyclic ring having five or six atoms in the ring;   X is oxygen or sulfur;   R 3  is a radical selected from the group consisting of hydrogen, C 1  -C 6  alkyl, hydroxy C 1  -C 6  alkyl, polyhydroxy C 1  -C 6  alkyl, phenyl, phenyl-C 1  -C 6  alkylene, C 1  -C 6  alkylphenyl, C 1  -C 6  alkoxyphenyl, halophenyl, trifluoromethylphenyl, hydroxy, oxo (O═), C 1  -C 6  alkoxy, phenyl-C 1  -C 6  alkoxy, halo, mercapto, thioxo (S═) , C 1  -C 6  alkylthio, phenyl-C 1  -C 6  alkylthio, C 1  -C 6  alkanoyl (C 1  -C 6  acyl), C 1  -C 6  alkoxy carbonyl, C 1  -C 6  alkylene C 1  -C 6  alkylcarboxylate, C 1  -C 6  alkoxy C 1  -C 6  alkyl carbonyl, and C 1  -C 6  alkyl carboxamido in which the carboxamido group has the formula CONR 9  R 10  wherein R 9  and R 10  are the same or different and are selected from the group consisting of hydrogen and C 1  -C 6  alkyl, or NR 9  R 10  together form a heterocyclic ring having five or six atoms in the ring;   R 4  is a beta-bonded aldoglycoside radical selected from the group consisting of 1'-aldopentosidyl, 1'-aldohexosidyl, mono-deoxygenated 1'-aldopentosidyl, and mono-deoxygenated 1'-aldohexosidyl, and their O-substituted C 1  -C 6  alkyl, C 1  -C 6  alkanoyl, benzyl, benzoyl and C 1  -C 6  acetal or ketal derivatives, an O-substituent other than an acetal or ketal, if present on one oxygen, being present on all available ring substituent oxygens;   the pharmaceutically acceptable salts of said agent; and   the tautomers thereof,   said immune response-enhancing agent being free from ionic charge in water at pH 7.2-7.4.   
     
     
       28. The process according to claim 27 wherein said immune response-enhancing agent has a structure that corresponds to a formula ##STR18## wherein Z is N--R 2  and ═X is ═O. 
     
     
       29. The process according to claim 28 wherein R 4  is 1'-aldopentosidyl in mono-deoxygenated 1'-aldopentosidyl. 
     
     
       30. The process according to claim 29 wherein R 1  is OH or SH. 
     
     
       31. The process according to claim 29 wherein R 2  is selected from the group consisting of C 1  -C 10  alkyl, C 3  -C 10  beta-alkenyl and halo C 1  -C 10  alkyl. 
     
     
       32. The process according to claim 29 wherein said immune response-enhancing agent is 7-allyl-8-oxoguanosine, 7-(1-chloroethyl)-8-oxoguanosine or 8-mercaptoguanosine.